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PubMed1980

Hydrocortisone suppression test and discriminant analysis in differential diagnosis of hypercalcaemia.

Experience is reported of the hydrocortisone suppression test in 140 hypercalcaemic patients, comprising 98 new cases of hyperparathyroidism and 42 cases of non-parathyroid malignant disease. The diagnostic accuracy of the test was compared in 168 patients with that of discriminant analysis, the discriminant functions being derived from plasma inorganic phosphate, alkaline phosphatase, chloride, bicarbonate, and urea, and the erythrocyte sedimentation rate. The hydrocortisone test and discriminant analysis each achieved a diagnostic accuracy of about 93% in 148 patients with either non-parathyroid malignant disease or hyperparathyroidism without osteitis fibrosa. When both tests pointed to the same diagnosis, they were wrong in less than 1% of cases. The hydrocortisone test was not helpful in patients with osteitis fibrosa. Both tests can be performed in any hospital with reliable standard laboratory services. Used in combination they have a high predictive value in distinguishing hypercalcaemia of parathyroid origin from that due to non-parathyroid malignant disease and have not led to errors of clinical importance. They should continue to play a major role in the differential diagnosis of hypercalcaemia until a prompt and reliable service finally establishes parathyroid hormone assay as the definitive laboratory procedure.

Adult↗
PubMed2006

Cloning and analysis of differentially expressed ESTs in swine muscle tissue.

The obvious difference in muscle growth and meat quality traits exists between Chinese indigenous pig and exotic pigs. In order to study the reason of these phenotypic differences and search the potential gene related to growth and meat quality traits, silver-stained mRNA differential display technique was used to detect the difference with mRNA of loin-eye muscle tissue from maturity pigs of Lantang in Guangdong Province and Large Yorkshire. One of the newly discovered expressed sequence tag (ESTsp3) was analyzed by using bioinformatic technique. The results showed: (1) nearly 2000 cDNA fragments were detected with 30 primer pairs, and 6 differentially expressed ESTs in the loin-eye muscle tissues from the two breeds were isolated and obtained. The differential fragments were cloned and sequenced. The all sequences were recorded in the GenBank. (2) The 786 bp fragment of ESTsp3 was obtained with in silico elongation system, the ORF analysis revealed that it existed as an 83 aa complete open reading frame, and the elongation sequences were verified by RT-PCR. The analysis of in silico expression profile showed that ESTsp3 is expressed in various growth stages and in most tissues and organs, such as soft tissue, skin, skeletal muscle and kidney, but with variant expression quantity.

Amino Acid Sequence↗
PubMed1997

Overdrive atrial stimulation during transesophageal electrophysiological study: usefulness of post-pacing VA interval analysis in differentiating supraventricular tachycardias with 1:1 atrio-ventricular relationship.

We evaluated the feasibility and usefulness of overdrive atrial pacing to identify the relationship between atrial and ventricular activation in supraventricular tachycardias with a stable 1:1 atrio-ventricular (AV) conduction ratio during a transesophageal electrophysiological investigation. Overdrive atrial stimulation was performed in 42 consecutive patients (11 males and 31 females; mean age 49 +/- 17 years) during AV junctional reentrant tachycardia, orthodromic AV reentrant tachycardia and ectopic atrial tachycardia (22, 13 and seven subjects, respectively). Trains of 12 stimuli at a constant rate were introduced starting at a cycle length 10 ms shorter than the tachycardia cycle length; stimulation was repeated with a 10-ms decrement in pacing cycle length at each step until tachycardia terminated and/or second-degree AV block occurred. The difference between the VA interval duration at baseline and in the first post-pacing tachycardia beat was measured at each step and provided identification of the AV relationship. At least one post-pacing VA interval was evaluable in 90% of the cases and measured 2 +/- 4 and 1 +/- 3 ms in AV junctional and AV reentrant tachycardia groups, respectively, and 83 +/- 42 ms in the ectopic atrial tachycardia group (P < 0.0000001 ectopic atrial tachycardia group vs. others). When three or more post-pacing VA intervals were obtained during the same tachycardia, a curve was constructed by plotting their values against the corresponding pacing cycle lengths. A curve could be constructed in 36% of the cases and was flat in all patients with AV junctional and AV reentry, while it was completely irregular in the ectopic atrial tachycardia group (P < 0.003). The analysis of post-pacing VA interval behaviour in response to overdrive atrial stimulation provides a rapid and reliable differentiation between supraventricular tachycardias with 1:1 AV conduction ratio during a transesophageal electrophysiological study.

Adult↗
PubMed2005

Combined affinity labelling and mass spectrometry analysis of differential cell surface protein expression in normal and prostate cancer cells.

Differences in the expression of cell surface proteins between a normal prostate epithelial (1542-NP2TX) and a prostate cancer cell line (1542-CP3TX) derived from the same patient were investigated. A combination of affinity chromatographic purification of biotin-tagged surface proteins with mass spectrometry analysis identified 26 integral membrane proteins and 14 peripheral surface proteins. The findings confirm earlier reports of altered expression in prostate cancer for several cell surface proteins, including ALCAM/CD166, the Ephrin type A receptor, EGFR and the prostaglandin F2 receptor regulatory protein. In addition, several novel findings of differential expression were made, including the voltage-dependent anion selective channel proteins Porin 1 and 2, ecto-5'-nucleotidase (CD73) and Scavenger receptor B1. Cell surface protein expression changed both qualitatively and quantitatively when the cells were grown in the presence of either or both interferon INFalpha and INFgamma. Costimulation with type I and II interferons had additive or synergistic effects on the membrane density of several, mainly peripherally attached surface proteins. Concerted upregulation of surface exposed antigens may be of benefit in immuno-adjuvant-based treatment of interferon-responsive prostate cancer. In conclusion, this study demonstrates that differences in the expression of membrane proteins between normal and prostate cancer cells are reproducibly detectable following vectorial labelling with biotin, and that detailed analysis of extracellular-induced surface changes can be achieved by combining surface-specific labelling with high-resolution two-dimensional gel electrophoresis and mass spectrometry.

Cell Line, Tumor↗
PubMed2006

Differential-display analysis of gene expression in livers from normal and partially hepatectomized mice.

Partial hepatectomy, resulting in the removal of approximately 70% of the liver, is widely utilized for studies of liver growth in experimental animals. The regenerative response is proportional to the amount of liver removed. Knowing when and where genes are expressed provides a strong clue as to its biological role. The RNA differential-display (DD) technique facilitates monitoring the differential expression of a large number of activated or suppressed genes under various biological conditions. To reveal mechanisms of liver regeneration, we performed a comparative analysis of gene expression during liver regeneration using DD. We sacrificed male Balb/c mice, aged 10 to 12 weeks, at 0, 24, 48, and 72 hours, and 1 and 2 weeks after PHx. The livers were weighed, and the amount of glutamic-oxaloacetate transaminase in serum measured. We extracted the total RNA from frozen liver tissue and confirmed the RNA quality using a lab-chip system. DD analysis was performed essentially as described by Liang and Pardee. Semiquantitative reverse-transcription polymerase chain reaction was performed to confirm the results of DD analysis. Of the 56 fragments that exhibited changed expression levels during PHx, 39 were cloned and sequenced. There were 31 known genes, 13 unknown genes, and 9 expressed-sequence tags. These results indicated that DD is a powerful approach for monitoring molecular events in the regenerating liver.

Animals↗
PubMed2003

Two-component systems in Prochlorococcus MED4: genomic analysis and differential expression under stress.

Two-component signal transduction systems, composed of histidine sensory kinases and response regulators, constitute a key element of the mechanism by which bacteria sense and acclimatize to changes in their environment. The availability of whole genome sequences permits a detailed analysis of these genes in cyanobacteria. In the present paper, we focus mainly on Prochlorococcus MED4, a strain adapted to surface oceanic conditions, for which six putative response regulators (rer) and five putative histidine kinases (hik) were identified. These numbers are comparable to those found in the other marine picocyanobacteria but much lower than those found in freshwater cyanobacteria. Moreover, the diversity of these genes is low in Prochlorococcus since most histidine kinases are related to a single group (type I) and most response regulators to a single family (OmpR). Under standard conditions, quantitative reverse transcription polymerase chain reaction revealed that one hik (hik03) and two rer (rer04 and rer05) genes were expressed at relatively high levels compared to the other two-component system genes. In response to high light exposure, a moderate increase (>5-fold) was observed in the expression of some putative rer genes (rer01, rer04, rer05, and rer06), whereas a smaller increase (<3-fold) in hik03 and hik04 mRNA levels was detected. In contrast, both cold and heat shocks decreased rather than increased the expression of most hik and rer genes.

Amino Acid Sequence↗
PubMed2005

Coherent expression chromosome cluster analysis reveals differential regulatory functions of amino-terminal and distal parathyroid hormone-related protein domains in prostate carcinoma.

Parathyroid hormone-related protein (PTHrP) has a number of cancer-related actions. While best known for causing hypercalcemia of malignancy, it also has effects on cancer cell growth, apoptosis, and angiogenesis. Studying the actions of PTHrP in human cancer is complicated because there are three isoforms and many derived peptides. Several peptides are biologically active at known or presumed cell surface receptors; in addition, the PTHrP-derived molecules can exert effects at the cell nucleus. To address this complexity, we studied gene expression in a DU 145 prostate cancer cell line that was stably transfected with control vector, PTHrP 1-173 and PTHrP 33-173. With this model, regulatory effects of the amino-terminal portion of PTHrP would result only from transduction with the full-length molecule, while effects pertaining to distal sequences would be evident with either construct. Analysis of the expression profiles by microarrays demonstrated nonoverlapping groups of differentially expressed genes. Amino-terminal PTHrP affected groups of genes involved in apoptosis, prostaglandin and sex steroid metabolism, cell-matrix interactions, and cell differentiation, while PTHrP 33-173 caused substantial increases in MHC class I antigen expression. This work demonstrates the distinct biological actions of the amino-terminus compared to distal mid-molecule or carboxy-terminal sequences of PTHrP in prostate carcinoma cells and provides targets for further study of the malignant process.

Journal Article↗
PubMed2006

[Analysis of differential expression genes related to different metastasis potential of adenoid cystic carcinoma using restriction fragments differential display PCR].

OBJECTIVE: To construct differential expression profiles of adenoid cystic carcinoma cell lines for screening candidate genes related to metastasis and to verify some candidate genes in adenoid cystic carcinoma. METHODS: Restriction fragments differential display PCR (RFDD-PCR) was used to set up gene expression profiles of adenoid cystic carcinoma cell lines-ACC-M and ACC-2, with high and low metastasis potential respectively. Candidate genes were screened through bioinformatics analysis. Then, a gene family of these candidate genes was checked using semi-quantitative reverse transcription-PCR(RT-PCR). RESULTS: Two gene expression profiles including 5420 gene fragments were constructed, 12 genes of a family called matrix metalloproteinase genes (MMPs) were observed obvious differentially expressed between two cell lines. Results of semi-quantitative RT-PCR also identified this different expression of MMP2,MMP7,MMP9,MMP14,MMP15 and MMP24. CONCLUSION: The construction of gene expression profiles of ACC-M and ACC-2 cell lines makes the foundation for seeking the target genes of adenoid cystic carcinoma. MMP2,MMP7,MMP9 and MMP15 may be relevant with carcinogenesis, development and metastasis of adenoid cystic carcinoma, and different metastasis potential may result from different subtype of MMPs gene family.

Carcinoma, Adenoid Cystic↗
PubMed2005

Validity of leukocyte count to predict the severity of acute appendicitis.

OBJECTIVE: To ascertain whether white blood cell (WBC) count with differential analysis may predict severity of disease in acute appendicitis. METHODS: We conducted this retrospective study on appendectomy patients from 1996 to 2001, at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia. We reviewed patient's age, gender, duration of symptoms, temperature on admission, WBC count including differential and the histological diagnosis of the appendicular specimen. We further analyzed the data of those patients found to have acute, gangrenous and perforated appendicitis to determine the correlation between a high WBC count and a more advanced form of appendicitis. RESULTS: Out of an aggregate of 232 patients, 162 were males and 70 females with a mean age of 23.7 years (range, 12-70 years). Mean duration of symptoms was 1.9 +/- 1.1 days, mean temperature 37.8 +/- 1.4 degrees C, with reported elevated WBC count in 167 (71.9%) and normal in 65 (28.1%) cases. Mean WBC counts in acute were 14.5 +/- 7.3 x 10(9)/L, gangrenous 17.1 +/- 3.9 x 10(9)/L and perforated appendicitis 17.9 +/- 2.1 x 10(9)/L. This reflected a persistently higher WBC count in the complex (gangrenous, perforated) appendicitis compared with acute appendicitis (p < 0.05). The differential analysis showed neutrophilia in 123 (53%) and lymphopenia in 112 (48%) cases and out of these, 116 (94%) with neutrophilia and 107 (95%) with lymphopenia were reported to have appendicitis. CONCLUSION: A high WBC with differential count is a reliable indicator of the severity of appendicitis and signifies a more advanced stage.

Acute Disease↗
PubMed1997

Molecular DNA analysis for differentiation of persistence or relapse from recurrence in treatment failure of Streptococcus pyogenes pharyngitis.

In the evaluation of treatment failure in group A streptococcal pharyngitis, it is essential to distinguish persistence or relapse with homologous streptococcal strains from the acquisition of new, unrelated strains. Randomly amplified polymorphic DNA analysis and restriction fragment length polymorphism analysis of total DNA were used as epidemiological tools to compare 122 pre- and post-treatment Streptococcus pyogenes isolates obtained from 61 patients. The results obtained by molecular typing showed that bacteriological failures were due to the original strains in 43 cases (70%) and to new strains in 18 cases (30%). In the present study, restriction fragment length polymorphism analysis of total DNA appeared to be more discriminative than randomly amplified polymorphic DNA analysis. Thus, molecular analysis of DNA is an effective way to distinguish recurrence from persistence or relapse and will be useful in assessing the efficacy of new antibiotic treatments.

Adolescent↗
PubMed2006

Alternative oxidases in Arabidopsis: a comparative analysis of differential expression in the gene family provides new insights into function of non-phosphorylating bypasses.

The emergence of Arabidopsis as a model plant provides an opportunity to gain insights into the role of the alternative oxidase that cannot be as readily achieved in other plant species. The analysis of extensive mRNA expression data indicates that all five Aox genes (Aox1a, 1b, 1c, 1d and 2) are expressed, but organ and developmental regulation are evident, suggesting regulatory specialisation of Aox gene members. The stress-induced nature of the alternative pathway in a variety of plants is further supported in Arabidopsis as Aox1a and Aox1d are amongst the most stress responsive genes amongst the hundreds of known genes encoding mitochondrial proteins. Analysis of genes co-expressed with Aoxs from studies of responses to various treatments altering mitochondrial functions and/or from plants with altered Aox levels reveals that: (i) this gene set encodes more functions outside the mitochondrion than functions in mitochondria, (ii) several pathways for induction exist and there is a difference in the magnitude of the induction in each pathway, (iii) the magnitude of induction may depend on the endogenous levels of Aox, and (iv) induction of Aox can be oxidative stress-dependent or -independent depending on the gene member and the tissue analysed. An overall role for Aox in re-programming cellular metabolism in response to the ever changing environment encountered by plants is proposed.

Arabidopsis↗
PubMedDate not supplied

Analysis of differential protein expression by cisplatin treatment in cervical carcinoma cells.

Cisplatin (cis-diaminedichloroplatinum), a DNA-damaging agent, which readily induces apoptosis in vitro, is one of the widely used anticancer drug in the treatment of human malignancies. Cisplatin has played an important role in cervical cancer management for effective chemotherapeutic regimen, but the underlying mechanisms inducing cell death at protein level are unknown. Using proteome analysis, an investigation aimed at a better understanding of the antiproliferative mechanisms by cisplatin was carried out in HeLa cervical carcinoma cells. In total, 21 protein spots were found to be differentially expressed following cisplatin treatment, of which 12 were upregulated (eg, regulator of G-protein signaling, TRAF:TNF (tumor necrosis factor) receptor-associated factor-interacting protein [I-TRAF], and cyclin-dependent kinase inhibitor p27 [p27(kip1)]) and 9 were downregulated (eg, myc proto-oncoprotein [c-myc] and proliferating cell nuclear antigen). Interestingly, we found the upregulation of proliferating cell nuclear antigen, which used molecular marker in cervical cancer screening. On the basis of proteomic data, we showed that cisplatin induced TRAF2-mediated NF-kappaB downregulation. In addition, our study demonstrated that cisplatin induced membrane death receptor-mediated and mitochondria-mediated apoptosis pathway. Our findings may offer new insights into the antiproliferative mechanism by cisplatin and its mode of action in cervical carcinoma cells.

Antineoplastic Agents↗
PubMed2003

Identification of Candida spp. by randomly amplified polymorphic DNA analysis and differentiation between Candida albicans and Candida dubliniensis by direct PCR methods.

Because Candida species have innately highly variable antifungal susceptibilities, the availability of a fast and reliable species identification test is very important so that suitable and effective therapeutic measures may be taken. Using three oligonucleotide primers, we established a randomly amplified polymorphic DNA (RAPD) analysis method that enabled direct identification of the most common opportunistic pathogenic Candida species. RAPD analysis revealed a characteristic molecular fingerprint for each Candida species. Differences between the profiles for Candida albicans and C. dubliniensis were evident. RAPD analysis is a relatively easy, reproducible, and reliable technique that can be useful in providing genetic fingerprints for the identification of strains. In addition, a collection of different C. albicans strains was identified by a specific PCR based on multiple secreted aspartic proteinase (SAP) genes and the dipeptidyl aminopeptidase (DAP2) gene. Our findings demonstrate that PCR based upon the SAP and DAP2 sequences is a simple, rapid, clear, and direct technique for the identification and differentiation of C. albicans and C. dubliniensis.

Candida↗
PubMed2026

damidBind: an R/bioconductor package for differential DamID analysis and data exploration.

SUMMARY: DamID, and its cell-type specific adaptations, including Targeted DamID (TaDa) and Chromatin Accessibility TaDa (CATaDa), are now widely-adopted as techniques for the genome-wide profiling of DNA binding proteins. Despite this popularity, no dedicated software solution exists for identifying differentially bound or accessible loci, or differentially transcribed genes, between cell types using DamID. The R/Bioconductor package damidBind provides these functions, allowing an end-user to move from processed binding profiles to identifying differentially-bound loci in a reproducible, statistically appropriate and straightforward workflow. AVAILABILITY AND IMPLEMENTATION: damidBind is an open-source R/Bioconductor package and freely available from Bioconductor at https://bioconductor.org/packages/damidBind/, and from GitHub at https://github.com/marshall-lab/damidBind. It is released under the GPLv3 licence.

Software↗
PubMed2003

Differential proteome analysis of TRAP-activated platelets: involvement of DOK-2 and phosphorylation of RGS proteins.

We have applied a proteomics approach to analyze signaling cascades in human platelets stimulated by thrombin receptor activating peptide (TRAP). By analyzing basal and TRAP-activated platelets using 2-dimensional gel electrophoresis (2-DE), we detected 62 differentially regulated protein features. From these, 41 could be identified by liquid chromatography-coupled tandem mass spectrometry (LC-MS/MS) and were found to derive from 31 different genes, 8 of which had not previously been reported in platelets, including the adapter downstream of tyrosine kinase 2 (Dok-2). Further studies revealed that the change in mobility of Dok-2 was brought about by tyrosine phosphorylation. Dok-2 tyrosine phosphorylation was also found to be involved in collagen receptor, glycoprotein VI (GPVI), signaling as well as in outside-in signaling through the major platelet integrin, alpha IIIb beta 3. These studies also provided the first demonstration of posttranslational modification of 2 regulator of G protein signaling (RGS) proteins, RGS10 and 18. Phosphorylation of RGS18 was mapped to Ser49 by MS/MS analysis. This study provides a new approach for the identification of novel signaling molecules in activated platelets, providing new insights into the mechanisms of platelet activation and building the basis for the development of therapeutic agents for thrombotic diseases.

Adaptor Proteins, Signal Transducing↗
PubMed1980

Evidence that (+)-bupranolol interacts directly with myocardial beta-adrenoceptors. Control of optical purity with differential thermal analysis.

Melting points measured with the capillary method were 150.5 degree C, 150.5 degree C and 224.0 degree C for hydrochlorides of (+)-bupranolol, (-)-bupranolol and (+/-)-bupranolol, respectively. The large difference in melting points of 73.5 degree C prompted us to determine possible contaminations of (+)-bupranolol with traces of (-)-bupranolol using differential scanning calorimetry. We detected as little as 0.001% (-)-bupranolol in a standard mixture of (+)-bupranolol and (-)-bupranolol. A batch of (+)-bupranolol not measurably contaminated with (-)-bupranolol (optically purity greater than 99.999%) was used in pharmacological and biochemical assays. The affinities of (-)-bupranolol and (+)-bupranolol were determined functionally by the blockade of isoprenaline stimulation of spontaneously beating rat right atria and electrically driven kitten papillary muscles; and directly by inhibition of binding of 3H-(-)-propranolol to kitten ventricle membrane particles. In all 3 systems the enantiomeric (-)/(+) affinity ratio was 50--120 for bupranolol. These experiments prove that (+)-bupranolol itself binds to the beta-adrenoceptors of mammalian myocardium.

Animals↗
PubMedDate not supplied

[Comparative analysis of differentiating and apoptogenic effects of cytidine, thymidine, and guanosine on cells of human erythromyeloleukemic cell line K562].

The antiproliferative, differentiating and apoptogenic effects of purinic (cytidine and thymidine) and pyrimidinic (guanosine) nucleosides on K562 cells were investigated. Induction of erythroid differentiation (haemoglobin synthesis) in cancer cells was observed after 2 and 4 days incubation with 2 mM thymidine and 1 mM guanosine. Increase of haemoglobin synthesis in K 562 cells incubated with 2 mM cytidine was detected at 4 day. Analysis of the DNA-degrading effect (induction of apoptosis) of the reagents used on K 562 cells by fluorescent dyes and modulation activity of caspase 9 and also caspases 3,7,10 is induced with thymidine and guanosine but not with cytidine.

Antineoplastic Agents↗
PubMed2005

cDNA microarray analysis of differential gene expression in Candida albicans biofilm exposed to farnesol.

Candida albicans biofilms are structured microbial communities with high levels of drug resistance. Farnesol, a quorum-sensing molecule that inhibits hyphal formation in C. albicans, has been found to prevent biofilm formation by C. albicans. There is limited information, however, about the molecular mechanism of farnesol against biofilm formation. We used cDNA microarray analysis to identify the changes in the gene expression profile of a C. albicans biofilm inhibited by farnesol. Confocal scanning laser microscopy was used to visualize and confirm normal and farnesol-inhibited biofilms. A total of 274 genes were identified as responsive, with 104 genes up-regulated and 170 genes down-regulated. Independent reverse transcription-PCR analysis was used to confirm the important changes detected by microarray analysis. In addition to hyphal formation-associated genes (e.g., TUP1, CRK1, and PDE2), a number of other genes with roles related to drug resistance (e.g., FCR1 and PDR16), cell wall maintenance (e.g., CHT2 and CHT3), and iron transport (e.g., FTR2) were responsive, as were several genes encoding heat shock proteins (e.g., HSP70, HSP90, HSP104, CaMSI3, and SSA2). Further study of these differentially regulated genes is warranted to evaluate how they may be involved in C. albicans biofilm formation. Consistent with the down-regulation of the cell surface hydrophobicity-associated gene (CSH1), the water-hydrocarbon two-phase assay showed a decrease in cell surface hydrophobicity in the farnesol-treated group compared to that in the control group. Our data provide new insight into the molecular mechanism of farnesol against C. albicans biofilm formation.

Biofilms↗